iXIGHIEM C D U CHI THf PHAIM TLT V I V E TIIMH (IVIICRaSATELLITE] D E CHOIM GIOIMG CA C H E P
[CYPRIIVUS CARPia^
Nguyin Hiru Ninh^
TOMTAT
Bai bao trinh bay kit qua tinh cac gia tri chpn loc tinh trang sinh tru'dng trong chuong trinh chpn gidng ca chep {Cyprinus carpio) sir dung chi thi phan tir vi ve tinh (microsatellite) xay dung pha he. Quan dan chpn gidng ban dau (GQ) dugc hinh thanh tir sau dan ca chep. Hai the he chpn gidng tiep theo gdm 135 gia dinh d the he Gj va 93 gia dinh d the he G,. Bay microsatellite da dugc sir dung de xac dinh huyet thdng, ty le thanh cdng xac dinh con theo bd me d the he G, la 96,8% (1284 ca the dugc xac dinh vao 113 gia dinh) va d the he Gj la 96,2% (1282 ca the dugc xac dinh vao 92 gia dinh). Phan tich di truyin cho kit qua ve he sd di truyen la 0,25 ddi vdi ca hai tinh trang trpng lupng va chieu dai d lln can do cudi ciing. Anh hudng cua di tmyIn theo me va cac yeu td mdi trudng rlt thap, gin bing khdng. Hieu qua chpn loc trung binh ddi vdi trpng luong la 9,9% va chieu dai la 5,2% d mdi the he chpn loc.
Tir khda: Ca chep, huyet thdng, tinh di truyen, dac diem sinh tnrdng.
I.DATVANDE
Ca chip II mot trong nhQng ddi tugng nuoi thuy san quan trpng tren t h i gidi, san lugng ciia e l c h i p udc tinh chiem 13% tdng san lugng nu6i trdng thuy san nude ngpt tren t h i gidi nam 2004 (FAO, 2004).
Cl chip cung II loli c l nuoi tmy In thdng d Viet Nam, cl dugc nuoi trong nhilu he thdng vdi hinh thirc nu6i khac nhau. Chuong trinh chgn gidng d c h i p d Viet Nam da dugc tiln hanh tir nam 1985 dya tren ba quIn din ban dau gdm e l c h i p trIng Viet Nam, ea chip vay Hungary v l e l c h i p vIng Ind6nexia. Sau nam thi he chgn loc theo phuong phip chpn Ipc c l the, tdc dd sinh trudng cua c l c h i p tang 33% so vdi quIn din ban diu (Thien, 1993). Phuong p h i p chgn gidng c l the cho thay hieu qua chpn lgc giam d the he thir nam, nen chuong trinh chgn gidng theo phuong phap chpn Ipc gia dinh duge thyc hien dya tren vat lieu di tmyIn ban diu II cle ddng c l chgn Ipc theo phuong phip chpn loc e l the. Sau hai t h i he chpn loc c l chgn Ipc gia dinh cd tdc dp sinh trudng tang 7% so voi quIn din ban dau. Tuy nhien, do quy m6 ciia chuong trinh chgn gidng ban hep, sd gia dinh dugc chgn lgc kh6ng nhilu nen han e h l d i n viec tilp tuc thuc hien chuong trinh chpn gidng.
Th6ng tin v l p h i he trong elc chuong trinh chpn gidng khong chi d n thilt cho viec tinh toln d c thong sd chpn loc m l cdn cho phip quIn ly vat lieu
Vien Nghien ciru Nuoi trong Thiiy san I
di tmyIn hieu q u i (Falconer va Mackay, 1996). Theo doi p h i he II mot trong nhung vIn d l khd khan nhat ddi vdi d c chuong trinh chgn gidng d c gidng loli thuy sin. The he eon eiia mdi gia dinh th6ng thudng phii dugc nudi rieng den khi dii ldn de cd the bin sd dien tir (PIT tag). Do dd, ylu cau sy dau tu ldn ve ca sd vlt chat v l nhan e6ng lao dong de nu6i rieng d e gia dinh; ngoli ra d c gia dinh nu6i rieng vIn ed nhiing tac dong d i n g ke cita cle yeu td m6i tmdng khIc nhau llm Inh hudng d i n viec tinh toan cac ehi sd di tmyIn chgn lgc (Herbinger va ddng tie gil, 1995).
Xlc dinh huyet thdng b i n g chi thi di tmyIn phan tii lln diu tien dugc Ip dung thanh c6ng tren e l hdi (Doyle v l Herbinger, 1994; Wright v l Bentzen, 1995). Hieu qua eua ky thuat nly da dugc kiem nghiem trong thi nghiem nudi chung e l hdi trout, k i t q u i da xlc dinh duge 91% d the theo d e gia dinh cua sa dd lai dly dii giua 10 c l dye vdi 10 c l d i , sir dung 4 chi thi phan tii microsatellite (Herbinger v l ddng tic gia, 1995). Sir dung 4 chi thi phan tu microsatellite (vi ve tinh) cho p h i p xlc dinh it nhat 99,5% cua tdng sd 792 d the eon theo gia dinh khi tiln hinh lai giua 100 c l dye voi 100 c l d i tren c l hdi Dai Tay Duong (O'Reilly vl ddng tie gia, 1998).
Chi thi phan tii microsatellite da dugc phlt trien cho c l c h i p (Crooijmans va ddng tac gia, 1997). Mue dich eiia nghien cim nly la sir dung chi thi phan fir microsatellite xle dinh huyet thdng ciia d c d the
nuoi chung ngay tu' giai doan ca bgt nhlm tinh toan d c th6ng sd di truyin chinh xlc hon, nang cao hieu qua chpn gidng sinh tm'dng e l c h i p tiln h i n h tai Vien Nghien cun Nu6i trdng Tliuy san I, Bic Ninh.
II. VAT UEU VA PHUDNG PHAP NGHIEN CUU
1. Vat Ueu di trayln v l tao quIn d i n ban diu GQ Vat lieu ban diu gdm slu quIn din c l chip: (1) ca chip chgn gidng gia dinh the he thir hai; (2) c l chip chpn gidng ca t h i dong Hungary; (3) c l c h i p vay Hungai-y; (4) ca c h i p chpn gidng ca thi ddng Indonexia; (5) c l chep vIng Indonexia; (6) ca c h i p chpn gidng c l the dong Viet Nam. Tdng sd 101 gia dinh da dugc tao ra theo sa dd lai d b a n g 1, mdi c l bd vl c l me chi tham gia san xult mot gia dinh. Sd lugng e l bd me cua moi ddng tham gia tao t h i he GQ II khIc nhau dya tren ca sd vl xep hang tdc do sinh tru'dng. Cac gia dinh dugc nuoi rieng trong giai d i n khi dat co 5-10 g/eon thi tiln hanh bin sd dien fir PIT tag, mdi gia dinh bin sd cho 25 c l the. Sau giai doan uong nuoi thi cd 15 gia dinh bl that tholt do ty le sdng thIp, cdn lai 86 gia dinh cd dii e l d l bin sd, d c ca the sau khi bin sd PIT tag da duge nu6i chung trong ciing mot ao.
Bang 1. So dd lai giira sau ddng ca tao quan din ban diu phye vu chgn gidng
Ca me 1 2 3 4 5 6
Ca bd 1
8(9) 2 9(9) 8(9)
3 7(7) 7(7) 6(7)
4 5(5) 4(5) 4(5) 5(5)
5 0(4) 1(4) 4(4) 3(3) 3(3)
6 3(3) 2(3) 1(3) 3(3) 3(3) 0(0)
* Ci bd vi ca me theo thu tu 1-6 li ten ciia 6 quin dan ci su dung tao quin din ban diu
* Cic sd trong ngoac kep li sd gia dinh sin xuit (101 gia dinh); sd ngoai ngoac kep li sd gia dinh cd du sdci the con debin sd (86 gia dinh).
2. The he chpn gidng thir nhat (Gi) vl thir hai {G^
CIc gia dinh chgn gidng d c l hai t h i he diu dugc Ip dung theo sa dd d bang 2, mdi c l bd va d me cung tham gia san xuat hai gia dinh. Sau khi tiem kich due td kich thich sinh sin vl c l d i rung tning thi vudt trung c l d i chia lam hai phln (30 g / p h i n ) , dung hai c l due khIc nhau de thu tinh vdi mdi phln trung de tao d c gia dinh full-sib (ddng huylt) vl half- sib (lai). Tdng sd 107 gia dinh da dugc san xult d t h i
he Gl tu' 58 ca bd va 58 ea me chgn loc tu' the he Go.
O t h i he G^, 77 gia dinh dugc sin xult fir 40 c l bd va 33 c l me chgn Ipc tu' 57 gia dinh cua t h i he Gj.
Tdng sd gia dinh ca ddi chung II 28 gia dinh a t h i he Gl vl 16 gia dinh d t h i he Gj da duge tao ra de so sinh voi d c gia dinh ca dupe chgn Ipc. Cac gia dinh ddi chung dugc tao ra khi tiln h i n h lai giua mot c l bd vdi mdt e l me.
Bang 2. Sa dd lai san xuat d e gia dinh chgn gidng d t h i he Gl vl G2 ( d c 6 b6i nIn dam la d c gia dinh
dugc san xuat tuong iing vdi e l bd v l me)
Ca nie
Vl F2 F3 F4 F5 F...
Fn
Ca bd
Ml M2 M3 M4 M5 M... Mn
3. Nu6i chung d e gia dinh v l thu sd Ueu sinh trudng d the he Gj va G2
Sau khi tning dugc thy tinh thi d c gia dinh dugc Ip rieng. Ngay sau khi ca bpt h i t noan hoIng thi chuyen 2000 con c l hot cua m6i gia dinh ra uong nu6i trong cung mot ao. Sau khi uong nu6i dugc 30 ngly thi giii lai nglu nhien 5000 c l the d l thi nudi tilp trong ao vdi mat dp 1 con/m^. Cl duge nuoi din khi bin sd PIT tag v l thu miu vay d l xlc dinh huylt thdng, ddng thdi can do chilu dii (cm) vl trpng lugng (g) lln thir nhat. Lln can do thii hai tiln hinh khoang 3 thing sau khi bin sd; lln can do cudi ciing tiln hinh khi d 12 thing tudi.
4. Xac dinh huyet thdng va theo doi pha he a. Thu miu vi tich chiet ADN
Khi lai de san xult d c gia dinh thudc the he chpn gidng thii nhat (Gi) vl thii hai (G2), c l bd me dupe ban sd PIT tag de theo doi va ddng thdi thu mau vay phuc vu xac dinh huylt thdng. Thu mau vay, b i n sd nglu nhien v l can do 1327 d the d thi he Gi v l 1332 c l the d t h i he Gj phuc vu phan tich pha he b i n g chi thi ADN. ADN duge tach chilt bing bp kit REAL(PieoPure).
b. Xac dinh sd lupng, chat lupng ADN vi sin phim PCR
ADN tach chilt tdng sd da duge xac dinh sd lugng vl chit lugng chinh xac tren may nanodrop
82 NONG NGHIEP VA PHAT TRIEN NONG THON - SO 4 - THANG 4/2010
ND-1000 spectrophotometer (Labtech International, UK). Ngoai ra, ky thuat dien di tren gel agaroza 1,5%
vdi dem lx SB (1 mM hydroxit natri, dung axit bone dilu chinh pH 8,5) dugc ap dung de kiem tra sd lupng, chit lupng cita ADN v l san phlm PCR.
c. Microsatellite vi tdi uu phin ung PCR
Mudi chi thi microsatellite da dugc lya chpn tir nghien ciiu cua Crooijmans va ddng tac gil (1997), trong dd chi cd 7 chi thi microsatellite cho k i t qua.
Hai phan img da thinh phln PCR da dugc phat triln d ba vi tri (triplex) cua microsatellite vl bdn vi tri (tetraplex) ciia microsatellite (Bang 3). Sau khi tdi un thi hai phln ung da thinh phln PCRs dugc tiln hinh chi tilt nhu sau: 15 pi phln ung gdm 60 ng ADN khuon, 150 pM mdi loai dNTPs, 1,5 mM MgCl,, lx dem PCR IV (75 mM Tris - HCl), dd HoO, 1 unit Taq polymeraza (gien AB) vl cle doan mdi (Chi tilt vl ndng dp d bang 2). Phln img PCR dugc tien h i n h budc diu tien biin tinh d 92°C trong 5 phiit, tiep theo la 35 chu ky gdm 40 giay d 92''C, 50 giay d 55'*C va 1 phut 45 giay d 72''C, chu ky cudi cung II 72°C trong 25 phut.
Bang 3. Hai phan img da thanh phan PCR xle dinh huyet thdng c l chip.
Multiplex 1
Multiplex 2
Ten vi tri microsate
llitc MF\V4 MFW9 MFWll MFW7 MFW18 MFW20 MFW26
Khoang alen (gien
diing vi) 102-166 79-194 132-240 181-285 86-297 125-252 88-165
Danh dau Blue Black Green Black Blue Green Black
Ndng dp mdi doan
mdi 0,10 pm/nl 0,25 pm/nl 0,14 pm/ul 0,16pm/nl 0,10pm/iil 0,22 pin/pl 0,14 pm/iil d. Phan tich alen vi xic dinh huyet thdng Cac alen dugc tich tren mIy giai trinh ty ty dong CEQ 8800. Dien di mao quan, mdi miu gdm 2 pi sin pham PCR, 28 pi formamit, 0,25 pi kich thuac tieu chuan (60 bp - 400 bp) va mot gipt diu. Phln m l m Genemaker 2.0 dugc sir dung de phan tich vl xlc dinh d c alen. Phln m l m VITASSIGN (Vandeputte vl ddng tic gil, 2006) dupe sir dung d l xlc dinh con theo bd me.
5. Phan tich elc thong sd di trayln
M6 hinh tiiyln tinh thing (GLM) phan tich anh hudng cua d c ylu td thi nghiem, phan tich tren phln m l m SAS (SAS Institute Inc., 1997). Tilp theo.
Mixed model bao gdm cac ylu td cd dinh nhu thi he.
dong ca, gioi tinh va tudi; d c ylu td nglu nhien gdm bd, me va tuong tac giu'a bd v l me dupe tap hgp trong m6 hinh phan tich PROC MIXED (Little va ddng tie gia, 1996). Chi tilt vl mo hinh Mixed model nhu sau:
Y.^w= Ii + GEN,+ LINE,-H SEX^, + AGE -i- S„-h D^ +1„_
H-e ijkiipq (Model 1)
Trong do, Y II gil tri quan sat cua e l the q, |i
ijkiipq
II gia trj tmng binh; GEN, II ylu td ed dinh cua t h i he (i= 1, 2, 3); LINE^ II ylu td cd dinh cua ddng c l (j
= 1, 2); SEX,i- II ylu td cd dinh eua gidi tinh {k=l,2)\
AGE la hiep phuong sai eua tudi; S„ la Inh hudng ngau nhien cua c l t h i bd thir n, D^ II Inh hudng ngau nhien cua c l t h i me thir p dam; I,,^^ la tuong tie giira c l t h i bd thir n vdi ca the me thir p, va eiji^,^^ II sai sd ngau nhien.
CIc gia tri phuong sai phan tich tir Model 1 dugc sir dung cho phan tich tilp theo tren phln mem ASREML (Gilmour va ddng tic gil, 2002). Phan tich bing Model 2, phan tich sir dyng quan he huylt thdng qua d c t h i he theo m6 hinh toln nhu sau:
Y,^,„= \i + GEN,+ LINE,+ SEX,.+AGE+1„+ D^+ e^^,„
(Model 2)
Trong dd, Y II gia tri quan sit cua c l t h i q, \i
ijknpq
II gia tri tmng binh; GEN, II ylu td cd dinh cua t h i he (/= 1, 2, 3); LINE^ II ylu td cd dinh ciia ddng c l (j
= 1, 2); SEX,(. II ylu td cd dinh cua gioi tinh {k= 1, 2);
AGE II hiep phuong sai cua tudi; I„ II Inh hudng di truyin cgng gdp ngau nhien ciia ca the thir n; D^ la anh hudng nglu nhien cua m6i trudng vl Inh hudng theo me; vl e^^..^,,, II sai sd nglu nhien.
Tinh mot sd th6ng sd di truyin: cr]^ II phuong sai theo me v l m6i trudng nencr,", = o]^^^:, de d l theo doi thi chung se duge sir dung II ol-.
Phuong sai kieu hinh {o],) dugc tinh dya tren phtrong sai do di trayln {<j\), phuong sai do m6i tru'dng {G]) va phuong sai do sai sd {u]) nhu sau:
o",",
Do dd, he sd di tmyen dupe tinh: Ir =
C6ng thire tinh anh hudng cua yeu td m6i trudng
2
vl theo me: c' =—^. Tuong quan kieu di tmyIn va cr;,
kieu hinh giua elc tinh trang dugc tinh:
trong dd CTI2 II phuong sai di truyin hoac kieu hinh giua hai tinh trang.
III. KET QUA NGHIEN CUU 1. Xac dinh huyet thdng
a. Mot sddac cfiem cua chi thi phin tu nucrosatelhte ddi vdi cic quin dan ci chep
CIc chi thi phan tir microsatellite nghien cim diu tuan theo quy luat phan ly Hardy-Weinberg. BIy microsatellite da tao ra tdng sd 107 alen d quin dan ea bin, sd alen d hai t h i he sau giam hon so vdi quin din ban diu.
Sd alen trung binh tren m6i vi tri microsatellite cung gilm tir 15,3 d t h i he GQ xudng 14,9 d t h i he Gi va 13,6 d t h i he G2. Ty le giam alen tir 1,6% a t h i he chpn gidng thir nhlt din 10,2% d t h i he chgn gidng thir hai so vdi quin din ban diu.
Bang 4. Da hinh cac microsatellite d the he GQ, GJ va G2 trong chuong trinh chgn gidng ea chip.
Bang 5. Hieu qua cua xac dinh huyet thdng sii dung bay ehi thi microsateUite qua hai the hp chgn lgc
T d n g sd alen T m n g binh sd
alen T m n g binh sd
alen hieu q u i
% alen bi m i t
Go 107 15,3 9,34 0
Gl
104 14,9 9,26 1,6
G2
95 13,6 9,03 10,2 b. Xic dinh huyet thdng d the he Gj vi G2 Kh6ng cd sy sai khIc dang k l v l ty le xlc dinh huylt thdng thinh cong giua hai t h i he. Sir dung 7 microsatellite xac dinh thinh c6ng huylt thdng cua con theo bd me II 96,8% d the he Gi vl 96,2% d t h i he G2. Kit q u i xlc dinh huyet thdng thinh c6ng tuang ty dat 91% tren e l hdi (Fishback vl ddng tic gil, 2002) vl 95% tren c l c h i p (Vandeputte vl ddng tie gil, 2004).
Ty le sd c l the dugc xlc dinh nhilu ban mot cap bd me tir 1,7% din 2,8%. Sd c l the con kh6ng dugc xlc dinh thugc gia dinh nIo II thIp d t h i he Gj (1,3%), d t h i he G2 k i t q u i cung tuong ty (1,2%).
Sy kh6ng hien dien cita con d i trong mot sd gia dinh cd the II do ty le sdng cita d c gia dinh thap, sd lupng miu it, phuong phip lay mau vl sai sd ti-ong qui trinh phan tich (O'Reilly v l ddng tic gia, 1998;
Herlin vl ddng tie gil, 2007; Castro vl ddng tac gia, 2006).
So gia dinh
Sd ca the con dugc phan tich xac dinh huyet thdng Ty le % thanh cdng xac dinh
con theo mdi cap bd me Ty le % ca the dugc xac dinh
nhieu hon mdt cap bd me Ty le % sd ca the con khdng
duoc xac dinh bd me Sd gia dinh khdng cd su xuat
hien ca the con nao Sd gia dinh dugc xac dinh cd
su xuat hien cua con cai
The he C;, 135 1327 96,8%
1,9%
1,3%
22 113
The he G, 93 1332 96,2%
2,6%
1,2%
1 92 c. Ddng gdp cua bd me vio quy md gia dinh 6 t h i he Gl (Hinh 1), 31,9% d c gia dinh cd tir 1 den 5 e l the con. Phln ldn d c gia dinh (khoang 43,4%) dugc xac dinh cd tir 6 din 15 c l t h i eon. Ty le kh6ng nhd (24,7%) d c gia dinh cd nhilu hon 15 ca the eon. O t h i he G2 (Hinh 2), ty le d c gia dinh cd tir 1 d i n 5 e l the eon II 24,2%. Cd 37,4% vl 29,3% d c gia dinh tuong ung cd tir 6 den 15 c l t h i eon vl 16 din 25 e l t h i con. Khoang 9,1% gia dinh cd nhilu hon 25 e l t h i con.
Hinh 1. Tan sd phan bd cua eon cai trong cac gia dinh d thi he G,.
3().()
2l)()
e 15,(1
e
fvin 11-15 ir>-2(l 21-25 >30 1 aiii(> *A:(iiJjxKkiJl
Hinh 2. Tan sd phan bd eiia con cai trong cac gia dinh d t h i he Gj.
84 NONG NGHIEP VA PHAT TRIEN NONG THON - SO 4 - THANG 4/2010
2. Phan tich di trayln sd lugng a. Thdng ke md ta ve miu phan tich
Sd lieu phan tich cac ylu td di truyin dugc tom tit d bing 7. Sd miu phan tich d cac lln cd khac nhau do sd lugng e l hi chit hoac m i t sd, ghi chep nhlm sd.
Bing 7. Sd miu, trang binh, do lech chuan, hiep phuong sai ddi voi trgng lugng va chilu dai cua c l d
ba lan can do d d c t h i he Gi v l G2
Hiep phuong sai cua tinh trang trpng lugng rat cao, fir 46,9% din 56,4%; trong khi dd chilu dii II tir 14,6-18,6%. Hiep phuong sai trong nghien cun nay cao hon so vol nghien cim cua Kocour va ddng tac gil (2007) vl tuong duong vdi nghien cun cua Wang vl ddng tic gil (2006). Dp lech chuan tang theo dp tudi vl ca eua cl. Do lech chuan rlt eao cho thIy cd sai khIc rlt ldn giua elc ca the trong cung quin din v l chilu dii vl trgng lugng.
b. Gii tri trung binh cua quin din vi cic the he Chilu dii vl trgng lugng cua c l thi nghiem ed sai khIe y nghia so vdi c l ddi chimg (P<0,05) d tit e l d c lln theo doi. Sai khIc cd y nghia (P<0,01) v l trgng lugng ciing quan sit duge khi so sinh giua hai t h i he d d c lln thu sd lieu.
Chilu dii ciia e l d t h i he Gj v l G2 khong ed sai khac y nghia d lln do thir nhlt vl thii hai, sai khac chi dat dugc d lln thu sd lieu thii ba. Sy sai khIc y nghia giua quin din chgn lgc so voi quin dan ddi chimg, hay sai khac giua elc t h i he cd the la do kit q u i ciia chpn Ipc (Bang 8).
Bang 8. Gil tri tmng binh nhd nhat (± sai sd chuan) eua d c tinh trang ddi vdi moi quan dan (chpn lgc va ddi chiing) va cae t h i he phan tich bang model h6n tap (Model 1).
Cac tinh trang Trpng lupng (g)
Chilu dai (cm)
Lln theo
doi 1 2 3 1 2 3
Sd miu 2555 2512 2391 2555 2512 2391
Trung binh 91,40 251,38 775,57 17,58 24,90 34,31
Dp lech chuln
45,5 118,0 437,0 2,7 3,6 6,4
Hiep phuong
sai (%) 49,8 46,9 56,4 15,2 14,6 18,6
Tinh trang Trpng lupng (g)
Chilu dii (cm)
Lln theo doi 1 2 3 1 2 3
Quin din Ddi chimg
73,5" ± 4,25 238,9" ± 8,83 687,7'- ± 25,95
15,9" ± 0,25 23,0^ ± 0,29 32,7" ± 0,33
Chpn Ipc 103,6''' ± 2,68 289,7'-' ± 5,02 763,2" ± 15,66
18,0'-' ± 0,16 25,3" ± 0,18 34,5'^ ± 0,19
T h i he G,
87,5" ± 7,49 226,9" ±11,0 661,9" ± 47,86
17,6" ± 0,45 25,2" ± 0,39 31,6" ± 0,83
G2 114,8'-'±8,18 252,4'-'±11,2 776,l'-y: 65,59
18,3" ± 0,48 25,5" ±0,37 36,6" ± 0,75
* Cac gia tri trung binh trong cimg nipt hang (ddi chung so vdi chpn loc; Gj so vdi GJ cd cac chir cai phia tren khac nhau thi sai khac cd y nghia (P<0,05).
c. Phuang sai va cac thdng sd di truyen
Phuong sai do sai sd ldn nhat quan sat dugc chiim 71,7% vl 74,4?^ tdng phuong sai cua trpng lupng vl chilu dii d lln can do cudi cung. Ty le phuong sai do elc ylu td moi tnrdng so vdi tdng phuong sai kieu hinh cao nhlt d lln can do thii nhat (3,8%khiblnsdPITtiig).
Anh hudng ciia cac ylu td moi tradng (c^ II rlt nhd, giam tii lln quan sit thu: nhlt (0,017 ± 0,006) din lln quan sit thu' ba (0,01 ± 0,003). He sd di truyin eiia trpng lugng vl chilu dai giam theo do
tudi cua cl, he sd di tmyen cua tinh trang sinh trudng ciia c l khi ban sd II 0,39, khi thu hoach II 0,25.
Kit qua dat dugc tuong ty nhu k i t qua nghiln cim cua Vandeputte v l ddng tie gia (2004), thIp hon so vdi nghien eiru cua Kocour v l ddng tie gia (2007).
Tuy nhien, hai nghien ciiu tren day kh6ng tinh toln anh hudng cua d c ylu td moi trudng, hon nira sd lugng gia dinh va sd e l the chgn loc la thap hon nhilu so vdi trong nghien cim nly.
Bang 9. Phuang sai do di tinyen cgng gop (a]), phuong sai do m6i trudng (o";"), phuang sai do sai sd (cr^"), he sd di trayln {h' ± sai sd chuan) va anh hudng eua m6i tmdng (c" ± sai sd chuan) cua trpng lugng va chilu dii bba lan
theo doi (Phan ta'eh bang Model 2).
CIc ylu td
1
<^A
•)
a-.
1
h'
1
c
Trpng lugng 1
813,81 78,59 1188,49 0,39 ± 0,06 0,038 ± 0,006
2 6214,60
140,72 12201,85 0,33 ± 0,05 0,008 ± 0,004
3 6477,20
880,62 18607,30 0,25 ± 0,04 0,034 ± 0,002
Chilu dai 1
3,43 0,15 5,19 0,39 ± 0,06 0,017 ± 0,006
2 5,93 0,24 12,42 0,32 ± 0,06 0,013 ± 0,004
3 8,36 0,35 25,38 0,25 ± 0,06 0,010 ± 0,003 d Tuong quan di truySn vi kieu hinh ^^n 0,021. Tuang quan di trayln vl kieu hinh giua
cle tinh trang sinh trudng tuong tu vdi d c kit qua Tuong quan di truyen giua hai tinh trang a mirc , . . - ,. - - , . - , .'• • ' i r j
^ ' , -' & *> nghien cun tren ca chep eua cac tac gia Vandeputte d6 trung binh den eao, tir 0,61 den 0,99 vdi sai s6 , ..i ., ., ^onn^\ wi • J - 4.- •' /OAA^\
• , , , va dong tac gia (2004), Wang va dong tac gia (2006), chuan thap. Tuong quan kieu hmh cua d c tinh trang „ , ,.; , ., /orvrvy-v
dao dong tii 0,58 din 0,99 vdi sai sd chuln tir 0,001
Bang 10. Tuong quan v l kieu hinh (niia tren dudng ehio eiia bang) v l di trayln (mia dudi dudng chIo cua bang) (± sai sd chuan) giira d e tinh trang
Wtl Lgi Wt2 Lg2 Wt3 Lg3
Wtl 0,97 ± 0,000 0,95 ± 0,023 0,94 ± 0,026 0,80 ± 0,100 0,91 ± 0,083
Lgi 0,99 ± 0,001 0,61 ± 0,065 0,97 ± 0,016 0,87 ± 0,088 0,99 ± 0,001
Wt2 0,90 ± 0,006 0,60 ± 0,002 0,85 ± 0,104 0,98 ± 0,016 0,93 ± 0,086
Lg2 0,87 ± 0,008 0,90 ± 0,006 0,93 ± 0,004 0,85 ± 0,094 0,99 ± 0,001
'Wt3 0,58 ± 0,019 0,58 ± 0,019 0,67 ± 0,014 0,64 ± 0,015 0,85 ± 0,087
Lg3 0,61 ± 0,021 0,62 ± 0,019 0,67 ± 0,016 0,73 ± 0,013 0,72 ±0,011
* Wtl, wt2, wt3; lgi, lg2. IgSli trpng lupng va chiSu e. Hieu qui ciia chpn Ipc
Hieu qui chpn Ipc dugc tinh theo gil tri tmng binh nhd nhlt cua ddng c l chpn Ipc so voi ddng ca ddi chimg theo model hdn tap. Hieu qua chpn Ipc ddi vdi trpng lugng cita e l dao dong tir 9,9-29,1% d mdi t h i he. Tinh trang ehilu dii ed hieu q u i chgn Ipc tir 5,2-11,7% d mdi t h i he.
IV. KET LUAN
Kit qui nghiln ciiu cho thay viec sir dung bay chi thi phan tir microsatellite cho p h i p xle dinh thinh cdng huylt thdng ciia cle quin din e l chip.
Nhung chi thi microsatellite nay cd the dugc sir dung de thilt lap pha he c l c h i p nu6i chung ngay tir giai doan c l hot mdi nd.
He sd di truyin cua tinh trang trpng lugng v l chieu dii e l chip II tu'ong duong nhau v l d mue trung binh (0,25) den cao (0,39). Hieu qua chpn Ipc cao khi sir dung d c th6ng tin pha he d l phan tich.
dii cua ci aba lin thu miu.
Phan tich tuong quan giua elc tinh trang cho thay viec chpn loc v l trpng lupng cung dem lai kit qua tdt v l chilu dii.
TAI UEU THAM KHAO
1. Castro, J., Pino, A., Hermida, M., Bouza, C, Riaza, A., Ferreiro, I., SInchez, L , Martinez, P., 2006.
A microsatellite marker tool for parentage analysis in Senegal sole (Solea senegalensis): Genotyping errors, null alleles and conformance to theoretical assumptions. Aquaculture 261, 1194-1203.
2. Crooijmans, R. P. M. A., Bierbooms, V. A. F., Komen, J., Vand Der Poel, J. J., Groenen, M. A. M., 1997. Microsatellite markers in common carp
(Cyprinus carpio L). Animal Genetic 28,129-134.
3. Doyle, R. W., and Herbinger, C. M., 1994.
Broodstock improvement strategies based on DNA fingerprinting-examples and cost-benefit analysis.
Aquaculture 137, 283.
86 NONG NGHIEP VA PHAT TRIEN NONG THON - SO 4 - THAMr:: A/onm
4. Falconer, D. S. and Mackay, T F. C , 1996.
Introduction to quantitative genetics. Longman, Essex CM20 2JE, England, 464 p.
5. FAO Fisheries Department, 2004. Fisheries Statistics.FAO-Rome.
http;//w\v\v,fag,grgZtigis/servlet/.
6. Fishback, A. G., Danzmann, R. G., Ferguson, M. M., Gibson, J. P., 2002. Estimates of genetic parameters and genotype by environment interactions for gro-wth traits of the rainbo'w trout {Oncorhynchus mykiss) as inferred using molecular pedigrees. Aquaculture 206, 137-150.
7. Gilmour, A R., Gogel, B. J., CuUis, B. R., Welham, S. J., Thompson, R., 2002. ASReml User Guide Release 1.0. VSN International Ltd, Hemel Hempstead, HPllES, UK.
8. Herbinger, C. M., Doyle, R. W., Pitman, E. R., Paquet, D., Mesa, K. A , Morris, D. B., Wright, J. M., and Cook, D., 1995. DNA fingerprint based analysis of paternal and maternal effects on offspring growth and survival in communally reared rainbow-trout.
Aquaculture 137, 245-256.
9. HerUn, M., Taggart, J. B., McAndrew, B. J., Penman, D. J., 2007. Parentage allocation in a complex situation: A large commercial Atlantic cod
(Gadus morhua) mass spawning tank. Aquaculture 274, 218-224.
10. Kocour, M., Mauger, S., Rodina, M., Gela, D., Linhart, O., Vandeputte, M., 2007. Heritiibnity estimates for processing and quality traits in common carp (Cyprinus carpio L.) using a molecular pedigree. Aquaculture 270,43-50.
11. Littell, R. C , Milliken, G. A., Stroup, W. W.
and Wolfinger, R. D., 1996. SAS system for mixed
12. O'Reilly, P. T , Herbinger, C , and Wright, J.
M., 1998. Analysis of parentage determination in Atlantic salmon (Salmo sala) using microsatellites.
Animal Genetics 29, 363-370.
13. SAS/STAT Software, 1997. Release 6.1. Gary, NC, U.S.A.
14. Thien, T M., 1993. A review of the fish breeding research and practices in Vietnam. In: K.L.
Main, E. Reynolds (eds.): Selective Breeding of Fishes in Asia and The United States. The Ocean Institute, Honolulu, 190-197.
15. Vandeputte, M., Kocour, M., Mauger, S., Dupont-Nivet, M., Guerry, D. D., Rodina, M., Gela, D., Vallod, D., Chevassus, B., Linhart, 0., 2004.
Heritability estimates for growth-related traits using microsatellite parentage assignment in juvenile common carp (Cyprinus carpio L ) . Aquaculture 235, 223-236.
16. Vandeputte, M., Mauger, S., Dupont-Nivet, M.
2006. An evaluation of allowing for mismatches as a way to manage genotyping errors in parentage assignment by exclusion. Molecular Ecology Notes 6, 265-267.
17. Wang, C , Li, S., Xiang, S., Wang, J., Liu, Z., Pang, Z., Duan, J., Xu, Z., 2006. Genetic parameter estimates for grovrth-related traits in Oujiang color common carp (Cyprinus carpio var. color).
Aquaculture 259,103-107.
18. Wright, J. M. and Bentzen, P., 1995.
Microsatellite: genetic markers for the future. In: G.
R. Carvalho and T. J. Pitcher (eds.): Molecular Genetics in Fisheries. Chapman & HaU, UK, 117-121.
models. Cary, NC: SAS Institute.
MICROSATELLFTE MARKERS IN GENETIC SELECTION OF COMMON CARP (Cyprinus carpio) Nguyen Huu Ninh Summary
The objective of this paper is to report estimates of genetic parameters for body traits selection in common carp (Cyprinus carpio) selective breeding programme in which pedigrees were determined by using microsatellite markers. The base population was synthesized following a single pair mating scheme from six carp stocks. The next two selected generations had 135 famihes in the G, and 93 families in the Gj.
Seven microsatellite loci were used for parentage assignment with 96.8% of the offspring (1284 individuals) and 96.2% offspring (1282 individuals) were unambiguously assigned to 113 families in the Gj and 92 families in the G, generations, respectively. The estimated heritability value of common carp was 0.25 for both weight and length at the final harvest. The overall obtained maternal and common environmental effects were consistently close to zero. The average of direct response to selection per generation was 9.9%
for body weight and 5.2% for length.
Key words: Common carp, paientage assignment, heritability, growth traits.
Ngudi phan bien: TS. La Tuan Nghia
